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Method for evaluating immune regulation effect of lactobacillus acidophilus

A technology of Lactobacillus acidophilus and immune regulation, which is applied in biochemical equipment and methods, microbial measurement/testing, material inspection products, etc. It can solve adverse inflammatory reactions, untestable, lack of cytokine and chemokine detection, etc. problems, to achieve the effect of eliminating the risk of disease

Inactive Publication Date: 2010-12-15
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These evaluation methods are only based on the positive or negative results of a certain detection index to judge whether it has an immunomodulatory effect, and have certain limitations for comprehensive, effective, and overall evaluation of the immune regulatory function; and lack of cytokines closely related to the immune regulatory function The detection of chemokines and chemokines cannot detect the adverse inflammatory response that it may have on the host

Method used

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  • Method for evaluating immune regulation effect of lactobacillus acidophilus
  • Method for evaluating immune regulation effect of lactobacillus acidophilus
  • Method for evaluating immune regulation effect of lactobacillus acidophilus

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Experimental program
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specific Embodiment approach 1

[0014] Specific embodiment one: the method for evaluating the immunomodulatory effect of Lactobacillus acidophilus in this embodiment is realized according to the following steps: One, the Lactobacillus acidophilus to be evaluated is cultivated to the late logarithmic growth period, discards the supernatant after centrifugation, and uses cold Wash with PBS (phosphate buffer saline) for 3 times, and then resuspend the bacteria with DMEM cell culture medium, and the bacterial concentration was 3×10 8 cfu / ml bacterial suspension; 2. Wash the polarized human colon adenocarcinoma cells Caco-2 twice with PBS, then add 1 mL of bacterial suspension as the experimental group, and add 1 ml of DMEM cell culture medium as the blank control group, at 37°C, 5% CO 2 Incubate together in the incubator for 2h; 3. Use the Trizol method to extract the total RNA of the human colon adenocarcinoma Caco-2 after incubation in the experimental group and the blank control group, and then extract the pr...

specific Embodiment approach 2

[0071] Embodiment 2: This embodiment differs from Embodiment 1 in that the centrifugation in step 1 is performed at a speed of 5000 r / min for 10 minutes. The steps and parameters are the same as those in the first embodiment.

specific Embodiment approach 3

[0072] Specific embodiment three: the difference between this embodiment and specific embodiment one is the culture process of the human colon adenocarcinoma cells Caco-2 in the polarized state in step two: a, cell growth medium for human colon adenocarcinoma cells Caco-2 Cultivate, change the medium every other day, and subculture for 5 days; b. Human colon adenocarcinoma Caco-2 cells in good growth state were mixed with 3×10 5 cells / cm 2 The initial amount was inoculated on a six-well cell culture plate at 37°C, 5% CO 2 The human colon adenocarcinoma cell Caco-2 in a polarized state was obtained by continuous culture in an incubator for 15 days; the cell growth medium in step a was composed of 90ml of DMEM cell culture medium, 10ml of fetal bovine serum, 1g of penicillin sodium and 1 g of streptomycin. The steps and parameters are the same as those in the first embodiment.

[0073] In this embodiment, the Caco-2 human colon adenocarcinoma cells in a polarized state can si...

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Abstract

The invention discloses a method for evaluating the immune regulation effect of lactobacillus acidophilus, and relates to a method for evaluating the immune regulation effect. The method comprises the following steps of: 1, culturing the lactobacillus acidophilus to be evaluated to a post-logarithmic growth phase to prepare bacterial suspension; 2, incubating human colonic adenocarcinoma cells Caco-2 in a polarized state and the bacterial suspension together; 3, extracting total RNA and protein from the incubated human colonic adenocarcinoma cells Caco-2 by a trizol method; and 4, detecting and comparing the expression level of an immune regulation related gene in the total RNA and the protein respectively by a real-time RT-PCR method and western blot method. The method can more objectively evaluate possible adverse inflammatory reaction and the like of the lactobacillus acidophilus on a host to eliminate potential pathogenic risk by detecting the immune regulation related gene.

Description

technical field [0001] The present invention relates to a method for evaluating immunomodulatory effects. Background technique [0002] Probiotic lactic acid bacteria are one of the most important physiological flora in the intestinal tract of humans and animals, and are closely related to various physiological functions of the body. Because of its many health benefits, its application in food and health products is increasing rapidly. The application of functional foods containing probiotics to promote health and prevent diseases has achieved many results, among which there are many reports on the role of probiotics in immunity. Although most probiotics, especially Lactobacillus "generally regarded as safe" (GRAS) strains, their possible pathogenic risk cannot be ruled out. [0003] The central role of the gut microbiota in normal immune responses suggests that altering and modulating the microbiota can have significant immunomodulatory effects. And studies have confirme...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/02G01N33/53
Inventor 姜毓君吕学娜王明娜韩琳琳张光辉满朝新刘颖曲行光
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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